Methods for reducing vein irritation

ABSTRACT

The present invention provides compositions for delivering highly water-soluble drugs (such as vinca alkaloids) and methods of using such compositions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/212,140, filed Aug. 17, 2011, allowed, which is a continuation ofU.S. application Ser. No. 12/965,676, filed on Dec. 10, 2010, now U.S.Pat. No. 8,026,250, which application is a continuation of U.S. patentapplication Ser. No. 10/889,226, filed on Jul. 12, 2004, now U.S. Pat.No. 7,871,632, all of which are incorporated herein by reference inentirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to an emulsion composition for delivering highlywater-soluble drugs such as vinca alkaloids.

2. Description of the Related Art

Despite years of research into the development of more effective andsafe therapeutic agents for cancer, cancer drugs remain extremely toxicto patients. The common systemic toxicities associated with cancerdrugs, such as chemotherapeutic agents, include bone marrow suppression,asthenia, peripheral neuropathy, dyspnea, alopecia, etc. To make thingsworse, many intravenously injected cancer drugs can cause localreactions at the injection site resulting in vein irritation, pain,tissue necrosis and/or thrombophlebitis. The consequence of injectionsite reactions to a chemotherapy agent may include extreme pain, earlytermination of drug treatment, wounds that are difficult to heal, scars,permanent tissue damage, and, in the worst case, amputation.

Alkaloids isolated from the periwinkle plant (Vinca rosea) andderivatives thereof, collectively referred to as “vinca alkaloids,” haveproven effective as first line therapy for many types of lymphomas,leukemias, and other cancers. Vincristine and vinblastine consist of acatharanthine moiety linked to vindoline, and the structures differ by asingle substitution in the vindoline group. Vindesine, a desacetylcarboxyamide derivative of vinblastine, was synthesized later.Subsequently, novel synthetic approaches were used to generate compoundsthat differed from the natural compounds by the presence of aneight-rather than a nine-member catharanthine ring, includingvinorelbine, which is commonly available as a tartrate salt, i.e.,vinorelbine bitartrate or vinorelbine tartrate.

Vinca alkaloids are highly cytotoxic drugs that disrupt microtubules,inhibit cell division and induce apoptosis. Without wishing to be boundto a particular theory, it is believed that vinca alkaloids exert theircytotoxic effects by binding to tubulin, the protein subunit ofmicrotubules.

Vincristine, vinblastine and vinorelbine are the best-known members ofthis drug family and are widely used clinically. Despite having similarstructures and mechanisms of action, vinca alkaloids differ in theirantitumor activity and toxicities. For example, vincristine is usedmostly to treat hematological cancer, and neurotoxicity is doselimiting. In contrast, vinorelbine is approved for the use as a singleagent to treat breast and non-small cell lung cancer, and its injectionsite reaction is most severe amongst all vinca alkaloid drugs.

It is well known that all vinca alkaloid drugs are associated withadverse reactions at the injection site. For example, the currentvinorelbine product approved in the U.S. (NAVELBINE®) has a “blackbox”warning due to its severe reaction at the injection site.

NAVELBINE® is reportedly associated with a high incidence (51%-61%) oflocal reactions at the injection site, characterized by injection sitepain and phlebitis. The injection site reaction or extravasation ofNAVELBINE® can be severe, ranging from considerable pain, irritation,and tissue necrosis to thrombophlebitis (The NAVELBINE® ProductInformation by GlaxoSmithKline).

NAVELBINE® (vinorelbine tartrate) is a simple solution formulation forintravenous administration. Each vial contains vinorelbine tartrateequivalent to 10 mg (1-mL vial) or 50 mg (5-mL vial) vinorelbine inwater for injection at pH 3.5.

Rittenberg reported post-incident care and management of venousirritation or phelibitis due to NAVELBINE® (Oncol. Nurs. Forum 22:707-10, 1995). Mare reported methods for preventing venous toxicity ofNAVELBINE® by co-administering anti-inflammatory drugs as defibrotide orketorolac, or changing infusion schedules from a bolus infusion to aslow infusion (Support Care Cancer 11: 593-6, 2003). However, theinjection site toxicity problem of vinorelbine or other vinca alkaloidshas not been properly addressed from the drug formulation approach andvinca alkaloid products remain the most venous toxic drugs.

Oil-in-water emulsion formulations may provide advantages over atraditional solution formulation such as the one used by NAVELBINE® incontrol of venous toxicity at injection site for irritating drugs. Forexample, the intramuscular or intravenous injection of erythromycin orclarithromycin in a solution formulation causes severe pain at theinjection site, and erythromycin or clarithromycin fat emulsion(oil-in-water) is locally non-irritating (WO 90/14094). However,oil-in-water emulsion formulations are typically applied to onlylipophilic drugs such as propofol, diazepam, erythromycin orclarithromycin, etc. Desai (U.S. Pat. No. 4,816,247) disclosed emulsioncompositions for administration of sparingly water soluble ionizablehydrophobic drugs.

Without wishing to be bound to a particular theory, it is believed thatin an oil-in-water emulsion, a lipophilic drug is preferentiallydissolved in the oil phase and therefore is coated and/or encapsulatedin the oil droplets, thus preventing direct contact of drug molecules ata high concentration with the venous endothelium tissue, thus reducingthe venous toxicity of the drug.

However, to date, the utility of an emulsion in preventing venoustoxicity of irritating drugs is limited to only lipophilic (orhydrophobic) drugs since highly water-soluble drugs, such as vincaalkaloid drugs, dol not partition well in the conventional emulsion oildroplets. For example, vinorelbine in the bitartrate salt form is highlysoluble in water with an aqueous solubility is >1000 mg/mL in distilledwater.

Thus, there remains a need in the art for developing compositions fordelivering highly water soluble drugs. The present invention fulfillssuch a need and provides other related advantages.

BRIEF SUMMARY OF THE INVENTION

The present invention provides compositions for delivering highly watersoluble drugs and methods for using such compositions.

In one aspect, the present invention provides compositions fordelivering highly water soluble drugs that comprises a triglyceride oil,an emulsifier, a stabilizer, and water, wherein the composition is anemulsion having an oil and an aqueous phase, and the drug issubstantially in the oil phase.

In certain embodiments, the highly water soluble drug is venous toxicand/or weakly basic absent the emulsion. In certain embodiments, thedrug is selected from the group consisting of dopamine, ciprofloxacin,vancomycin, norvancomycin, doxorubicin, daunorubicin, vinca alkaloids(e.g., vinorelbine), and pharmaceutically acceptable salts thereof.

In certain embodiments, the triglyceride oil is a triglyceride havinglong chain fatty acids, a triglyceride having medium chain fatty acids,or a mixture thereof.

In certain embodiments, the emulsifier is egg lecithin, soy lecithin, asynthetic phospholipid, or a mixture thereof.

In certain embodiments, the stabilizer is a fatty acid (e.g., oleicacid), riboflavin-5-phosphate, vitamin-E succinate, cholesterol sulfate,or a mixture thereof.

In certain embodiments, the drug has an aqueous solubility of at leastor over 30, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450,500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg/ml.

In certain embodiments, the charge ratio of the drug and the stabilizeris 1:1 to 1:10. In certain embodiments, the charge ratio of the drug andthe stabilizer is within the range of 5:1 to 3:1, 2:1 to 1:1, or 1.5:1to 1:1.

In certain embodiments, no less than 80%, 85%, 90%, 92%, 94%, 95%, 96%,97%, 98%, or 99% of the drug is present in the oil phase of theemulsion.

In certain embodiments, the drug in the emulsion composition is in aconcentration range of about 1 to about 50 mg/ml. In certainembodiments, the concentration of the drug in the emulsion is about 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mg/ml.

In another aspect, the present invention provides a composition fordelivering a highly water soluble, venous toxic and weakly basic drugthat comprises an oil-in-solid dispersion prepared by freeze-drying theemulsion described herein.

In another aspect, the present invention provides a lyophilizedformulation of a highly water soluble, venous toxic and weakly basicdrug, wherein the formulation, when hydrated, produces the emulsiondescribed herein.

In certain embodiments, when the lyophilized formulation isreconstituted in a liquid medium to provide particles, the particlesincrease in size by less than one-fold as compared to particles beforelyophilization.

In another aspect, the present invention provides a method for treatingcancer comprising administering to a patient in need thereof thecompositions described herein wherein the drug in the composition is avinca alkaloid (e.g., vinorelbine bitartrate). In certain embodiments,the composition is administered intravenously. In certain otherembodiments, the composition is administered intramuscularly orintraarterially.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the effects of various stabilizers on vinorelbineincorporation into the oil phase of an emulsion.

FIG. 2 shows the effects of concentrations of sodium oleate on theincorporation of vinorelbine into an emulsion.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compositions for delivering highly watersoluble drugs (including those that are also venous toxic and/or weaklybasic). The pharmaceutical compositions are oil-in-water emulsionscontaining sub-micron size oil droplets, and comprise triglyceride oil,stabilizers, emulsifiers, and water. The compositions may optionallycomprise preservatives and/or other inactive ingredients.

The term of “highly water-soluble drugs,” as used herein, refers to adrug (in its freebase or salt form) having solubility in water in excessof 30 mg/ml at room temperature (20-25° C.). The solubility of a drugmay be described in a variety of ways. The USP/NF generally expressesthe solubility in terms of the volume of solvent required to dissolve 1gram of the drug at a specified temperature (e.g., 1 g aspirin in 300 mlH₂O, 5 ml ethanol at 25° C.). Other references may use more subjectiveterms to describe solubility, such as those given in the following tablefrom Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa., latest edition.

Parts of solvent needed Descriptive terms for 1 part solute Very soluble   <1 Freely soluble 1-10 Soluble 10-30  Sparingly soluble 30-100Slightly soluble 100-1000 Very slightly soluble  1000-10,000 Practicallyinsoluble or insoluble >10,000

Therefore, the “highly water-soluble drugs” of this invention includethe drugs in the top 3 solubility categories, i.e., “very soluble,”“freely soluble,” and “soluble.”

The term of “venous toxic drugs,” as used herein, refers to drugs which,when intravenously injected in a solution formulation, can cause localreactions at the injection site that result in vein irritation, pain,tissue necrosis and/or thrombophlebitis.

The term “weakly basic drugs,” as used herein, refers to drugs having atleast one weakly basic functional group.

A “highly water-soluble, venous toxic and weakly basic drug” is commonlyprovided in a salt form. Examples of some commercial drugs, which fallinto this category include, but are not limited to, dopaminehydrochloride, ciprofloxacin lactate, vancomycin hydrochloride,norvancomycin hydrochloride, doxorubicin hydrochloride, daunorubicinhydrochloride, vincristine sulfate, vindestin sulfate, vinblastinesulfate, and vinorelbine bitartrate.

In certain embodiments, the highly water-soluble drugs for use in thisinvention are anti-neoplastic agents.

In other embodiments, the highly water-soluble drugs for use in thisinvention are vinca alkaloids, and the pharmaceutically acceptable saltsand derivatives thereof. Vinca alkaloids include, but are not limitedto, vincristine, vinblastine, vindesine and vinorelbine.

In yet further embodiments, the highly water soluble drug for use inthis invention is vinorelbine, and the pharmaceutically acceptable saltsthereof.

Pharmaceutically acceptable salts,” as used herein, refers to thosesalts which are, within the scope of sound medical judgment, suitablefor use in contact with the tissues of humans and lower animals withoutundue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use in the chemotherapy and prophylaxis of cancers. Amongthe more common pharmaceutically acceptable salts of vinca alkaloids arethe tartrate, sulfate and hydrochloride forms. Other acid salts used inthe pharmaceutical arts include adipate, acetate, bromide, mesylate,lactate, succinate, maleate, lactobionate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, gluconate, glycerophosphate, heptonate, hexanoate,hydrobromide, hydroiodide, 2-hydroxy ethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate,pantothenate, pectinate, persulfate, 3-phenylpropionate, picrate,pivalate, propionate, thiocyanate, tosylate, and undecanoate.

In certain embodiments, the vinorelbine salt is the bitartrate salthaving a chemical name is3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate (1:2) (salt)].

As used herein, the term “oil-in-water emulsion” refers to a colloidaldispersion systems in which liquid oil is dispersed in small droplets(the discrete phase) in an aqueous medium (the continuous phase),wherein in excess of 80% of the drug is dissolved and remains in the oildroplets. In certain embodiments, greater than 85%, 90%, 92%, 94%, 95%,96%, 97%, 98%, or 99% of the drug is present in the oil phase:

As used herein, the term “an acidic pH” is meant the pH measured for theemulsion to be in the range of pH of 2 to 6. In certain embodiment, theemulsion has a pH of 3 to 5, pH 3 to 4, or pH 3.5 to 4.0.

As used herein, the term “sub-micron size droplet” is meant oil dropletsin an oil-in-water emulsion having an average diameter of less than 1micron as measured by conventional sizing techniques such as laser lightscattering spectrometry. In certain embodiments, the oil droplets of thecompositions of the present invention have an average diameter of lessthan 500, 450, 400, 350, 300, or 250 nm. Oil droplets of sub-micron sizeare desired for the safe passage of these droplets in the capillaryblood vessel in the circulation. Droplets of greater than 5 micron indiameter are believed to be unsafe for intravenous injection since theymay block the capillary vessel resulting in pulmonary embolism. Incertain embodiments, the oil droplets of the compositions of the presentinvention have an average diameter of less than 0.2-micron (200 nm) sothe emulsion may be sterilized by filtering through a 0.2 micron sizedfilter membrane.

In certain embodiments, the oil droplets of the compositions of thepresent invention have an average diameter of less than about 150, 100,75, 50, 25, 20, 15, or 10 nm.

“Triglyceride oil,” as used herein, refers to a triglyceride compositionwhich is liquid at room temperature (20-25° C.), and which comprisesprimarily triglycerides of C6 to C18 fatty acids. Triglyceride oil isused in this invention to form the discrete phase of the emulsion or theoil droplets in which the drug is encapsulated. The triglyceride oil isthus desired to be non-toxic, biocompatible, and capable of formingstable droplets of the desired size and encapsulating the highlywater-soluble drugs.

The triglyceride oil used in this invention can be glycerol esters ofshort chain (C4 to C6), medium chain (C8-C12), or long chain (C14 toC18) fatty acids or mixture thereof.

In certain embodiments, triglyceride oils of medium chain fatty acidsmay be used. Such oils comprise predominantly glycerol triesters of C8to C12 fatty acids. These oils can be prepared synthetically bywell-known techniques, or can be obtained from natural sources by knowntechniques of thermal or solvent fractionation of suitable natural oils,such as palm oil or coconut oil, to yield fractions rich in the desiredlow-melting triglycerides. An exemplary low-melting, low molecularweight triglyceride oil is a low molecular weight fraction of coconut orpalm oil which is rich in mixed esters of caprylic (octanoic) and capric(decanoic) acids. Such oil is commercially available as Miglyol 812 fromSASOL GmbH Germany, CRODAMOL GTCC-PN from Croda Inc. of Parsippany,N.J., or Neobees M-5 oil from PVO International, Inc., of Boonton, N.J.Other low-melting medium chain oils may also be used in the presentinvention.

In certain embodiments, triglyceride oils with a high percentage ofglycerol triesters of unsaturated or polyunsaturated C14 to C18 fattyacids (long chain fatty acids) may be used. An example of such an oil issoybean oil, which typically has a fatty acid composition of about 80%oleic and linoleic acids. An injectable grade of soybean oil iscommercially available as Super Refined USP grade oil from Croda Inc. ofParsippany, N.J. Another example of such an oil is safflower oil. Otherlow-melting vegetable oils or low-melting fractions of oils, includingcottonseed, menhaden, olive, peanut, corn, sesame and flaxseed oil,which can be obtained by conventional thermal or solvent fractionation,may also be used in the present invention. While such unsaturated orpolyunsaturated vegetable oils may offer a cost advantage in formulatingcompositions according to this invention, they also exhibit a greatertendency to oxidative deterioration, and may require the addition of oilsoluble antioxidants, such as tocopherols.

The triglyceride oil is generally present in a range of from about 2 toabout 40% in the final emulsion formulation. In certain embodiments,triglyceride oil is present at about 5%, 10%, 15%, 20%, 25%, 30%, or 35%by weight in the final emulsion formulation.

In certain embodiments, the triglyceride oil comprises a 1:1 weightratio mixture of a medium chain triglyceride and a long chaintriglyceride.

As used herein, the term “stabilizers” refers to those ingredients thatretain the highly water-soluble drug in the oil droplets of anoil-in-water emulsion.

In other embodiments, the “stabilizers” comprise compounds selected fromgroups of fatty acids, cholesterol sulfate, riboflavin-5-phosphate, andvitamin E succinate or a mixture thereof.

Exemplary fatty acids include saturated fatty acids, monoenoic acids andpolyenoic acids. The saturated fatty acids include, but are not limitedto, those listed in the table below:

Shorthand Molecular Systematic name Common name designation wt. butanoicButyric  4:0 88.1 pentanoic Valeric  5:0 102.1 hexanoic Caproic  6:0116.1 octanoic Caprylic  8:0 144.2 nonanoic Pelargonic  9:0 158.2decanoic Capric 10:0 172.3 dodecanoic Lauric 12:0 200.3 tetradecanoicMyristic 14:0 228.4 hexadecanoic Palmitic 16:0 256.4 heptadecanoicmargaric (daturic) 17:0 270.4 octadecanoic Stearic 18:0 284.4 eicosanoicArachidic 20:0 412.5 docosanoic Behenic 22:0 340.5 tetracosanoicLignoceric 24:0 368.6 hexacosanoic Cerotic 26:0 396.7 heptacosanoicCarboceric 27:0 410.7 octacosanoic Montanic 28:0 424.8 triacontanoicMelissic 30:0 452.9 dotriacontanoic Lacceroic 32:0 481 tritriacontanoicceromelissic (psyllic) 33:0 495 tetratriacontanoic Geddic 34:0 509.1pentatriacontanoic Ceroplastic 35:0 523.1

Exemplary monoenoic fatty acids include those listed in the table below:

Shorthand Molecular Systematic name Common name designation wt.cis-4-decenoic Obtusilic 10:1(n-6) 170.3 cis-9-decenoic Caproleic10:1(n-1) 170.3 cis-5-lauroleic Lauroleic 12:1(n-7) 198.4cis-4-dodecenoic Linderic 12:1(n-8) 198.4 cis-9-tetradecenoicmyristoleic 14:1(n-5) 226.4 cis-5-tetradecenoic Physeteric 14:1(n-9)226.4 cis-4-tetradecenoic Tsuzuic 14:1(n-10) 226.4 cis-9-hexadecenoicpalmitoleic 16:1(n-7) 254.4 cis-6-octadecenoic petroselinic 18:1(n-12)282.4 cis-9-octadecenoic oleic 18:1(n-9) 282.4 cis-11-octadecenoicvaccenic (asclepic) 18:1(n-7) 282.4 cis-9-eicosenoic Gadoleic 20:1(n-11)310.5 cis-11-eicosenoic Gondoic 20:1(n-9) 310.5 cis-11-docosenoicCetoleic 22:1(n-11) 338.6 cis-13-docosenoic Erucic 22:1(n-9) 338.6cis-15-tetracosenoic Nervonic 24:1(n-9) 366.6

Exemplary polyenoic fatty acids include those listed in the table below:

Shorthand Molecular Systematic name Common name designation wt.9,12-octadecadienoic linoleic 18:2(n-6) 280.4 6,9,12-octadecatrienoiclinolenic 18:3(n-6) 278.4 8,11,14-eicosatrienoic dihomolinolenic20:3(n-6) 306.5 5,8,11,14-eicosatetraenoic arachidonic 20:4(n-6) 304.57,10,13,16-docosatetraenoic — 22:4(n-6) 332.6 4,7,10,13,16- — 22:5(n-6)330.6 docosapentaenoic 9,12,15-octadecatrienoic — 18:3(n-3) 278.46,9,12,15- stearidonic 18:4(n-3) 276.4 octadecatetraenoic8,11,14,17-eicosatetraenoic — 20:4(n-3) 304.5 5,8,11,14,17- EPA20:5(n-3) 302.5 eicosapentaenoic 7,10,13,16,19- DPA 22:5(n-3) 330.6docosapentaenoic 4,7,10,13,16,19- DHA 22:6(n-3) 328.6 docosahexaenoic5,8,11-eicosatrienoic Mead acid 20:3(n-9) 306.5

In certain embodiments, the fatty acid used is oleic acid.

The amount of stabilizers in the emulsions of this invention may be, bycharge ratio to the highly water-soluble drug, within a range of about5:1 to about 3:1, about 2:1 to about 1:1, or 1.5:1 to 1:1.

As used herein, the term “emulsifiers” refers to compounds that allowthe formation of a stable oil-in-water emulsion wherein the droplets areof sub-micron size and contain the highly water-soluble drug. Exemplaryemulsifiers include compounds selected from phospholipids, bile salts,polyoxylene sorbitan fatty acid esters (e.g., TWEENS), polyoxyethylenecastor oil derivatives (e.g., CREMOPHOR), albumin and poloxamer (e.g.,PLURONIC), and mixtures thereof.

A “stable oil-in-water emulsion” refers to an oil-in-water emulsionwherein more than 50% of the oil droplets in the emulsion do notincrease their size more than one-fold under appropriate storageconditions for at least 3 months.

In certain embodiments, the emulsion of the present invention is stablefor at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or 24months.

In certain embodiments, the average size of the oil droplets in theemulsion of the present invention does not increase by about 10%, 20%,30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 140%, 160%, 180%, 200%,or 250%.

In further embodiments, phospholipids may be used as emulsifiers.Phospholipids are available from naturally occurring sources or byorganic synthesis. Lecithin is a naturally occurring mixture of thediglycerides of stearic, palmitic, and oleic acids, linked to thecholine ester of phosphoric acid, commonly called phosphatidylcholine.Hydrogenated lecithin is the product of controlled hydrogenation oflecithin.

According to the United State Pharmacopoeia (USP), lecithin is anon-proprietary name describing a complex mixture of acetone-insolublephospholipids, which consists primarily of phosphotidylcholine,phosphotidylethanolamine, phosphotidylserine and phosphotidylinositol,combined with various amounts of other substances such as triglycerides,fatty acids, and carbohydrates.

Pharmaceutically, lecithins are mainly used as dispersing, emulsifying,and stabilizing agents and are included in intramuscular and intravenousinjections, parenteral nutritional formulations and topical products.Lecithin is also listed in the FDA Inactive Ingredients Guide for use ininhalations, intramuscular and intravenous injections, oral capsules,suspensions and tablets, rectal, topical, and vaginal preparations.

Phospholipids can also be synthesized and the common syntheticphospholipids are listed below:

Diacylglycerols

-   1,2-Dilauroyl-sn-glycerol (DLG)-   1,2-Dimyristoyl-sn-glycerol (DMG)-   1,2-Dipalmitoyl-sn-glycerol (DPG)-   1,2-Distearoyl-sn-glycerol (DSG)

Phosphatidic Acids

-   1,2-Dimyristoyl-sn-glycero-3-phosphatidic acid, sodium salt    (DMPA,Na)-   1,2-Dipalmitoyl-sn-glycero-3-phosphatidic acid, sodium salt    (DPPA,Na)-   1,2-Distearoyl-sn-glycero-3-phosphatidic acid, sodium salt (DSPA,Na)

Phosphocholines

-   1,2-Dilauroyl-sn-glycero-3-phosphocholine (DLPC)-   1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC)-   1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)-   1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)-   1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC)-   1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC)

Phosphoethanolamines

-   1,2-Dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE)-   1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)-   1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE)-   1,2-Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)

Phosphoglycerols

-   1,2-Dilauroyl-sn-glycero-3-phosphoglycerol, sodium salt (DLPG)-   1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol, sodium salt (DMPG)-   1,2-Dimyristoyl-sn-glycero-3-phospho-sn-1-glycerol, ammonium salt    (DMP-sn-1-G,NH4)-   1,2-Dipalmitoyl-sn-glycero-3-phosphoglycerol, sodium salt (DPPG,Na)-   1,2-Distearoyl-sn-glycero-3-phosphoglycerol, sodium salt (DSPG,Na)-   1,2-Distearoyl-sn-glycero-3-phospho-sn-1-glycerol, sodium salt    (DSP-sn-1G,Na)

Phosphoserines

-   1,2-Dipalmitoyl-sn-glycero-3-phospho-L-serine, sodium salt (DPPS,Na)

Mixed Chain Phospholipids

-   1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)-   1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol, sodium salt    (POPG,Na)-   1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol, ammonium salt    (POPG,NH4)

Lysophospholipids

-   1-Palmitoyl-2-lyso-sn-glycero-3-phosphocholine (P-lyso-PC)-   1-Stearoyl-2-lyso-sn-glycero-3-phosphocholine (S-Iyso-PC)

Pegylated Phospholipids

-   N-(Carbonyl-methoxypolyethyleneglycol 2000)-MPEG-2000-DPPE-   1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, sodium salt-   N-(Carbonyl-methoxypolyethyleneglycol 5000)-MPEG-5000-DSPE-   1,2-distearoyl-sn-glycero-3-phosphoethanolamine, sodium salt-   N-(Carbonyl-methoxypolyethyleneglycol 5000)-MPEG-5000-DPPE-   1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, sodium salt-   N-(Carbonyl-methoxypolyethyleneglycol 750)-MPEG-750-DSPE-   1,2-distearoyl-sn-glycero-3-phosphoethanolamine, sodium salt-   N-(Carbonyl-methoxypolyethyleneglycol 2000)-MPEG-2000-DSPE-   1,2-distearoyl-sn-glycero-3-phosphoethanolamine, sodium salt

In certain embodiments, the phospholipids in the formulations of thepresent invention may be egg or soy lecithin.

The amount of phospholipids, by weight, in the emulsions of the presentinvention may be within a range of about 2% to about 15%, such as atabout 5% to about 10%.

As used herein, the term “bile salts” refers to salts of bile acid,i.e., steroids having 1-3 hydroxyl groups and a five-carbon atom sidechain terminating in a carboxyl group, which can be conjugated toglycine or taurine. Bile salts include, but are not limited to, cholate,deoxycholate, chenodeoxycholate, or ursodeoxycholate, and their glycineor taurine conjugates, e.g., glycodeoxycholate (GDC), glycocholate (GC),or taurodeoxycholate (TDC).

As used herein, the term “preservatives” refers to agents that canprevent microbial growth in the emulsion formulation of this invention.The oil-in-water emulsions of this invention contain nutrients formicrobes and may thus be conducive to microbial growth or contamination.Therefore, a preservative may be desirable in the formulation,especially for a vialed product that is intended to provide multipledoses where multiple punctures of the vial stopper by syringe needlesare needed. The preservatives useful for this invention include, but arenot limited to, sodium edetate (EDTA), sodium metabisulfite, sodiumbenzoate, benzyl alcohol, bronopol, parabens, cresol, phenol,phenoxyethanol, phenylmercuric acetate, phenylmercuric borate,phenylmercuric nitrate, sorbate, benzoate, sorbic acid thimerosal,cetrimide, chlorhexidine, chlorobutanol, chlorocresol, benzalkoniumchloride and benzethonium chloride or a mixture thereof.

The other inactive ingredients used in the emulsion compositions of thisinvention may include virtually any component, such as an acid or basefor pH adjustment such as hydrochloric acid and sodium hydroxide, agentsto adjust the tonicity of the emulsion including sodium chloride,mannitol, sucrose, dextrose, lactose, polyethylene glycols (PEG) andglycerin or a mixture thereof.

The emulsion formulation of the present invention can be prepared sothat it is ready-to-use or can be prepared with a cryoprotectant(s) as alyophilized solid, i.e., “an oil-in-solid dispersion system” that can bereconstituted at a later date and diluted with water to reform theoil-in-water emulsion before injection.

As used herein, the term “an oil-in-solid dispersion system” refers to asolid matrix prepared by freeze-drying (lyophilizing) an oil-in-wateremulsion of this invention and the solid matrix can reform anoil-in-water emulsion of similar droplet size upon mixing with water(reconstitution). In certain embodiments, the average droplet size ofthe reformed emulsion is no more than about 500%, 300%, or 150% of theaverage droplet size of the emulsion before the freeze-drying. Anoil-in-solid dispersion system of this invention may be optionallyprepared by spray drying.

“Cryoprotectants” used in the emulsion compositions of the presentinvention refers to those ingredients which are added to maintain thediscrete and submicron droplets of the emulsion during the freeze-dryingprocess and, upon the removal of aqueous phase of the emulsion, toprovide a solid matrix for the droplets to form the an oil-in-soliddispersion system.

Cryoprotectants that may be used in the emulsion compositions of thisinvention include, but are not limited to, polyols, monosaccharides,disaccharides, polysaccharides, amino acids, peptides, proteins, andhydrophilic polymers, or mixtures thereof.

Polyols that may be used in the present invention include, but are notlimited to, glycerin, mannitol, erythritol, maltitol, xylitol, sorbitol,polyglycitol or mixtures thereof.

Monosaccharides that may be used in this invention include, but are notlimited to, glucose, mannose, fructose, lactulose, allose, altrose,gulose, idose, galactose, talose, ribose, arabinose, xylose, lyxose ormixtures thereof.

Disaccharides that may be used in this invention include, but are notlimited to, sucrose, lactose, maltose, isomaltose, trehalose, cellubioseor mixtures thereof.

Polysaccharides that may be used in this invention include, but are notlimited to, cellulose, amylose, inulin, chitin, chitosan, amylopectin,glycogen, pectin, hyaruronic acid or mixtures thereof.

Amino acids that may be used in this invention include, but are notlimited to, alanine, arginine, asparagine, aspartic acid, cysteine,glutamine, glutamic acid, glycine, histidine, isoleucine, leucine,lysine, methionine, phenylalanine, proline, serine, threonine,tryptophan, tyrosine, valine or mixtures thereof.

Peptides that may be used in this invention include, but are not limitedto, diglycine and triglycine.

Proteins that may be used in this invention include, but are not limitedto, albumin, collagen, casein, and gelatin.

Hydrophilic polymers that may be used in this invention include, but arenot limited to, polyethylene glycols povidones, poloxamers, polyvinylalcohols or mixtures thereof. The most preferred hydrophilic polymersare polyethylene glycols and povidones.

The concentration of the cryoprotectants used in the liquid emulsioncompositions may be in the range of about 2% to about 40% w/w, such asabout 5% to about 20% w/w and about 10% to about 15% w/w.

Generally, the emulsion formulation of the present invention can beprepared by performing one or more of the following steps:

-   -   (1) Dissolve the drug, triglyceride oil, emulsifier(s), and        stabilizer(s) in a sufficient amount of a volatile solvent, such        as ethanol, methylene chloride, chloroform, isopropanol,        methanol, tert-butylalcohol, etc. to form a clear solution,    -   (2) Remove the solvent to a toxicologically acceptable residual        level by vacuum or by blowing solution with a nitrogen or air        stream to obtain an oil phase.    -   (3) Dissolve the preversative(s), cryoprotectant(s), and other        inactive ingredients in water to form an aqueous phase.    -   (4) Optionally filter the oil and/or aqueous phase to remove        particles.    -   (5) Add the aqueous phase to the oil phase, and mix well to form        a crude oil-in-water emulsion.    -   (6) Adjust pH of the crude emulsion to the desired pH range.    -   (7) Pass the crude emulsion through a high pressure homogenizer,        such as a Microfluidizer 110F equipped with an emulsion        interaction chamber by Microfluidics Corp, MA, operating at        approximately 18000 psi pressure for 2 to 10 passages until the        emulsion droplets reaches the desired average size range and the        emulsion is free of droplets of greater than 5 microns in        diameter.    -   (8) Aseptically pass the emulsion through a sterile 0.2        micron-rated membrane filter to sterilize the emulsion.    -   (9) Aseptically fill the filtered emulsion into appropriate        sterile containers and seal the containers with appropriate        sterile stoppers.    -   (10) Optionally the emulsions may be freeze-dried to form the        oil-in-solid dispersion system.    -   (11) Perform necessary tests on the final emulsion or the        oil-in-solid dispersion system.

The compositions of the present invention may be administered to ananimal in need thereof via various routes, such as intravenous,intramuscular, intra-articular, intra-peritoneal, or oraladministration.

The present invention also provides methods for using the compositionsdescribed herein. For instance, the present invention provides methodsfor treating cancer that comprise administering to a patient in needthereof compositions that comprise highly water soluble anti-neoplasticdrugs (e.g., vinorelbine bitartrate).

The following examples are intended to illustrate the invention withoutlimiting the practice thereof.

EXAMPLES Example 1 pH-Stability Profiling of Vinorelbine in Selected pHBuffers

This study was to determine the range of pH in which vinorelbine is moststable. Vinorelbine solutions at 97.0 μg/mL concentrations were preparedat pH 1.95, 3.05, 5.98, 7.01 and 8.04 in sodium phosphate buffers, andat pH 4.01 and 5.01 in sodium acetate buffers.

The buffered venorelbine solutions were stored at 40° C. and analyzed byHPLC for vinorelbine concentrations at various time points. The recoveryof vinorelbine over the initial concentration, representing thestability of vinorelbine in each buffer is shown in Table 1. The pHrange in which vinorelbine is most stable was defined based on themaximum recovery.

TABLE 1 PH STABILITY STUDY OF VINORELBINE AT 97.0 μG/ML AT 40° C./75%RELATIVE HUMIDITY (RH) -% RECOVERY OVER 0 DAY TIME POINT Time point pHpH pH pH pH pH pH (day) 1.95 3.04 4.01 5.01 5.98 7.01 8.04 3 97.9 99.998.2 97.5 97.9 96.1 92.1 7 97.3 99.7 100.4 98.6 105.0 94.8 88.0 14 90.298.1 97.0 97.6 96.9 91.4 85.1

Conclusion: vinorelbine appeared most stable at pH between 3 and 5, thisacidic pH range was thus chosen for the emulsion formulations used inthe other examples.

Example 2 Effect of Stabilizers on Vinorelbine Incorporation into TheOil Phase of an Emulsion

Because of the high solubility of venorelbine in water, itsincorporation in a normal oil phase was found minimal. In other words,without a stabilizer, venorelbine is present primarily in the aqueousphase.

The purpose of this study was to determine the effect of selectedstabilizers on vinorelbine incorporation into the oil phase of anemulsion. Since venorelbine is a weak base, a stabilizer is preferred tobe an acid with lipophilic property. Four stabilizers including sodiumoleate, vitamin E succinate, riboflavin-5-phosphate sodium andcholesterol sulfate were evaluated. Each stabilizer contains an acidgroup head and a lipophilic tail, and is generally consideredappropriate for injection.

Emulsions used for this study contained 1.4% (w/w) vinorelbinebitartrate, 10% (w/w) soybean oil, 1.2% (w/w) soy lecithin(Phospholipon® 90G by PHOSPHOLIPID GmbH), 0.005% (w/w) disodium EDTA,and 2.25% (w/w) glycerol. The concentration of each stabilizer addedinto formulation is: 1.57% (w/w) for sodium oleate, 2.73% (w/w) forvitamin E succinate, 2.46% (w/w) for riboflavin-5-phosphate sodium and2.52% (w/w) for cholesterol sulfate. A formulation without anystabilizer (“control”) and a formulation with extra Phospholipon 90Gwere also evaluated in the study.

The drug incorporation into the oil phase was tested using a dialysismethod developed specifically for this purpose. This method involvedfilling 500 mg of an emulsion into a Slide-A-Lyzer dialysis cassettewith 10,000 MW cutoffs, placing the cassette in 70 mL phosphate bufferedsaline (PBS), pH 7.4 and shaking the solution on a platform shaker at100 RPM. A small volume (1 mL) of PBS was removed at each time point andanalyzed by HPLC for vinorelbine concentration. The time profiles ofvinorelbine concentration in PBS are shown in FIG. 1. The stabilizerthat provided a high incorporation and small, uniform and stableemulsion droplets was selected as the preferred stabilizer for theemulsion formulations used in other examples.

Conclusion: The selected sterilizers were able to maintain vinorelbinein the oil droplets as demonstrated by significant reduction in thevinorelbine concentration in PBS, since the droplets are incapable ofpassing the dialysis membrane due to their size. Vitamin E succinate,sodium oleate and riboflavin-5-phosphate appeared to be the mosteffective stabilizers.

Example 3 Effect of Concentration of Sodium Oleate on the Incorporationof Vinorelbine into Emulsion

Having determined sodium oleate being a strong stabilizer, emulsions ofthe same composition as in Example 2 were prepared with sodium oleate at1.57% (w/w), 1.18% (w/w), 0.78% (w/w) or 0.39% (w/w), which correspondto a vinorelbine-to-oleate molar ratio of 1:4, 1:3, 1:2 or 1:1, or acharge ratio of 1:1, 1:0.75, 1:0.5 or 1:0.25, respectively. A dialysistest was performed under the same conditions as described in Example 2,and eight time points were taken for twenty-eight hours. PBS sampleswere analyzed by HPLC for vinorelbine concentration in phosphatebuffered saline at each time point. The result is shown in FIG. 2. Theoptimal concentration of sodium oleate was determined based on theincorporation result and emulsion stability.

Conclusion: Sodium oleate added at a vinorelbine-to-oleate molar ratioof 1:4 (charge ratio of 1:1) almost completely stopped the partition ofvinorelbine into the aqueous phase.

Example 4 Stability of Vinorelbine in Emulsions at a Neutral pH

Having determined the preferred stabilizer and stabilizer concentration,a preliminary emulsion was prepared and tested for short-term stability.The purpose of this study was to determine the possibility of a pHneutral emulsion based on vinorelbine stability. The emulsion preparedcontained the following composition:

% (w/w) Vinorelbine bitartarate 1.389 (equivalent to 10 mg/mLvinorelbine freebase) Soybean oil 10 Soy lecithin 1.2 Disodium EDTA0.005 dihydrate Glycerol 2.25 Sodium oleate 1.57 Water QS to 100 pH 7.4

Both physical and chemical stability of the emulsion were tested in thisexperiment. The physical stability was examined by measuring the averagedroplet size using laser light scattering spectrometry (Malvernzetasizer 3000) and by observing the gross and microscopic appearance ofthe emulsion. The chemical stability is represented by the vinorelbinestability and was examined by HPLC analysis.

TABLE 2 CHEMICAL STABILITY OF THE EMULSION AT PH 7.4 Time point Storage% Drug recovered (day) condition over time 0 7 −20° C.   100.3 25° C.92.4 40° C. 90.9 18 −20° C.   98.8 25° C. 93.0 40° C. 89.6

TABLE 3 PHYSICAL STABILITY OF THE EMULSION AT PH 7.4 ≧5 micron Timedroplets by point Storage AVG droplet microscopic (day) conditionAppearance diameter (nm) examination 0 Uniform 179.2 ± 1.3 None 7 −20°C.    7889.7 ± 3682.5 Many  5° C. Not uniform, 178.5 ± 1.3 None someprecipitate 25° C. Not uniform, 180.5 ± 2.0 None some precipitate 40° C.Not uniform, 189.2 ± 2.2 None some precipitate 18 −20° C.   Phaseseparation  2695.5 ± 2056.5 Many  5° C. Not uniform, 179.9 ± 0.4 Somesome precipitate 25° C. Uniform 185.1 ± 2.8 Many 40° C. Uniform 196.0 ±0.6 Many

Conclusion: Vinorelbine emulsion at pH 7.4 is not stable both physicallyand chemically.

Example 5 Effect of Emulsion pH on Emulsion Uniformity and Droplet Size

After the pH-neutral emulsion (Example 4) failed to show satisfactorychemical stability, this study was performed to examine stability ofemulsions at various pH (Example 1).

In addition, medium chain triglyceride (MCT) was added to replace 50% ofthe soybean oil. A brand product of MCT (Miglyol® 812 by Sasol) wasused. The addition of MCT was to improve the emulsion stability andreduce the droplet size. The combination of soybean oil and MCT at 1:1ratio has been used in a number of IV fat emulsion product and thus wasconsidered acceptable as a carrier for venorelbine.

The emulsion formulation contained:

% (w/w) Venorelbine 1.39 Soybean oil 5 Miglyol 812N 5 Soy lecithin 1.2Disodium EDTA 0.005 dihydrate Glycerol 2.25 Sodium oleate 1.57 Deionizedwater QS

pH of the emulsions was adjusted to 7.03, 5.97, 4.97, 4.52, 3.95 and3.74 using 0.5N HCI. The physical stability of these emulsions wasexamined and their appearance is summarized in Table 4. An emulsion withthe most uniform in appearance and smallest and most stable droplet sizewas selected.

TABLE 4 PHYSICAL APPEARANCE OF EMULSIONS AT VARIOUS PH EmulsionFilterability through 0.2 AVG droplet pH micron filters diameter (nm)Appearance 7.03 Easy 169.8 ± 2.8 Uniform 5.97 Didn't go through 0.2 μmNA Not uniform 4.97 Easy 420.4 ± 8.3 Not uniform 4.52 Easy 471.9 ± 30.9Not uniform 3.95 Easy 220.0 ± 1.0 Not uniform 3.74 Easy 191.1 ± 1.6Uniform

Conclusion: Vinorelbine emulsions of uniform appearance and smalldroplet size (<200 nm) were only possible at pH 7.0 or pH 3.7.

Example 6 pH 3.7 Emulsion Stability

A portion of the pH 3.7 emulsion prepared in Example 5 was used tomonitor the physical and chemical stability of the emulsion. Theemulsion was aliquoted into 2 mL glass vial and stored at −20° C., 25°C. and 40° C. Up to 4-week stability results were generated (Tables 5and 6).

TABLE 5 PHYSICAL STABILITY OF THE PH 3.74 EMULSION AVG Time Dropletpoint Storage diameter (day) condition (nm) Appearance Microscopicappearance 7 −20° C. 121.4 ± 2.7 Uniform Uniform, No ≧5 micron droplets  25° C. 115.2 ± 0.7 Uniform Uniform, No ≧5 micron droplets   40° C.123.6 ± 8.5 Uniform Uniform, Some ≧5 micron droplets 14 −20 149.0 ± 0.3Uniform Uniform, Some ≧5 micron droplets   25° C. 112.8 ± 1.3 UniformUniform, No ≧5 micron droplets   40° C. 145.2 ± 2.6 Uniform Uniform,Some ≧5 micron droplets 28 −20° C. NA Not uniform NA   25° C. 115.3 ±5.3 Uniform Uniform, No ≧5 micron droplets   40° C. 113.2 ± 2.4 UniformUniform, some small particles

TABLE 6 CHEMICAL STABILITY OF THE PH 3.75 EMULSION Vinor- Time elbineVinor- RRT: 1.07-1.25 point Storage recovered elbine % % (day) condition(%) Purity (%) Impurity#1 % Impurity#2 7 −20° C. 98.4 0.93 0.92   25° C.101.8 98.0 0.68 1.64   40° C. 99.8 97.9 1.21 1.14 14 −20 98.5 0.74 0.68  25° C. 101.1 98.5 0.69 0.67   40° C. 98.2 97.8 0.91 1.06 28 −20° C.97.8 0.91 1.78   25° C. 98.5 98.1 1.03 0.79   40° C. 95.7 96.6 1.92 1.99

Conclusion: The pH 3.7 vinorelbine emulsion appeared physically andchemically stable at 25° C.

Example 7 Dilution Study

This study was performed to determine the method by which the emulsionshould be diluted for intravenous administration. Since the preferableemulsion is acidic, a neutralization and/or dilution step was tested forpurpose of intravenous infusion.

The pH 3.7 liquid emulsion was diluted to 5.0 mg/mL vinorelbine freebasewith 53 mM arginine freebase or 50 mM sodium hydroxide at a 1:1 volumeratio. The pH of the diluted/neutralized emulsion was 7.08 with 53 mMarginine and was 7.29 with 50 mM sodium hydroxide. The emulsions werefurther diluted to 0.5 mg/mL and 2 mg/mL vinorelbine freebase with 5%dextrose solution (D5W) for droplets stability monitoring at roomtemperature.

In another study, the pH 3.7 emulsion was diluted with D5W or lactatedringer's (LR) injection without arginine or sodium hydroxide asneutralizing agent. Again, the diluted emulsions were evaluated fordroplets stability. The stability results of diluted emulsion are shownin Tables 7 and 8.

TABLE 7 EMULSION DROPLET SIZE UPON NEUTRALIZATION AND DILUTION Dilutedwith Sample Neutralize D5W to ZAve (nm) at ZAve (nm) ID agent (mg/ml) pHtime 0 at 7 hour #1 NaOH 0.5 8.05 150.3 ± 5.1 181.4 ± 32.0 #2 NaOH 2.07.70 138.9 ± 18.5 120.6 ± 3.5 #3 Arginine 0.5 7.36 136.8 ± 5.9 212.6 ±6.6 #4 Arginine 2.0 7.29 138.9 ± 18.4 130.8 ± 10.0 #5 NA Diluted with5.05 151.1 ± 10.8 265.5 ± 21.0 LR to 0.5 #6 NA Diluted with 4.68 136.7 ±1.2 179.2 ± 26.7 LR to 2.0 #7 NA Diluted with 4.11 124.3 ± 2.2 188.8 ±0.9 D5W to 0.5 #8 NA Diluted with 3.73 185.4 ± 14.1 147.4 ± 10.3 D5W to2.0

TABLE 8 EMULSION APPEARANCE UPON NEUTRALIZATION AND DILUTION Dilutedwith Microscope Microscope Sample Neutralize D5W to check check at IDagent (mg/ml) pH at time 0 7 hour #1 NaOH 0.5 8.05 Some Some ≧5 micron≧5 micron droplets droplets #2 NaOH 2.0 7.70 Some Some ≧5 micron ≧5micron droplets droplets #3 Arginine 0.5 7.36 Some Some ≧5 micron ≧5micron droplets droplets #4 Arginine 2.0 7.29 Some Some ≧5 micron ≧5micron droplets droplets #5 NA Diluted with 5.05 No ≧5 micron Some LR to0.5 droplets ≧5 micron droplets #6 NA Diluted with 4.68 Some Some LR to2.0 ≧5 micron ≧5 micron droplets droplets #7 NA Diluted with 4.11 No ≧5micron No D5W to 0.5 droplets ≧5 micron droplets #8 NA Diluted with 3.73No ≧5 micron No D5W to 2.0 droplets ≧5 micron droplets

Conclusion: The pH 3.7 vinorelbine emulsion of Example 5 may be dilutedwith D5W prior to intravenous infusion.

Example 8 Preparation of Vinorelbine Emulsion for Stability, VeinIrritation And Acute Toxicity Study

In this example, an emulsion with the following composition wasproduced:

% (w/w) Vinorelbine bitartrate 1.4 Miglyol 812N 15 Soy lecithin 7.5Disodium EDTA dihydrate 0.005 Oleic acid 1.5 Sucrose 15 Deionized waterto QS 100 HCl to adjust pH to 3.5 +/− 0.2

The batch size was 108 mL. The following describes the method ofpreparation:

-   -   A. An oil phase was prepared by dissolving vinorelbine tartrate,        Miglyol 812N, Soy lecithin, and oleic acid in a sufficient        quantity of dehydrated ethanol to form a clear solution. The        ethanol was removed using a Rotavapor (BÜCHI R-114) to a        residual ethanol concentration of <1%.    -   B. An aqueous phase was prepared by dissolving sucrose and        disodium EDTA dihydrate in water for Injection.    -   C. The oil and aqueous phases were mixed together using a        Silverson homogenizer (Model L4RT with a 2″ head) at        5,000-10,000 RPM for about 5 minutes to form a crude emulsion.    -   D. The pH of this crude emulsion was adjusted from 3.5 using 1N        HCl.    -   E. The crude emulsion was homogenized for six passes in the        Microfluidizer Model 110S.    -   F. In a laminar flow hood, the emulsion was filtered through a        0.45 μm filter and then a 0.2 μm sterile filter (Sartorius,        MiniSart).    -   G. The filtered emulsion was dispensed in 5 mL aliquots into 5        mL pre-sterilized glass vials. These vials were sealed with        pre-sterilized rubber stoppers.

Example 9 Stability of Vinorelbine Emulsion

The chemical stability of vinorelbine in the emulsion prepared inExample 8 was studied using reverse-phase HPLC method. This methodallows determination of concentration and purity of vinorelbine in theemulsion. The vinorelbine chemical stability data of the emulsion areshown in the table below:

Chemical stability Time Storage Conc. point Condition (μg/mL) % Recovery% Purity 0 NA 14.0 100.0 98.5 Wk 1 −20° C. 14.4 100.0 98.4 2-8° C. 14.6101.2 98.7 25° C. 14.3 98.9 98.9 40° C. 14.5 100.7 98.6 Wk 2 −20° C.14.3 100.0 99.0 2-8° C. 13.9 97.5 99.3 25° C. 14.1 98.6 98.9 40° C. 14.299.5 98.5 Wk 4 −20° C. 14.8 100.0 99.0 2-8° C. 14.7 99.4 99.1 25° C.14.4 97.3 98.7 40° C. 14.6 98.5 97.2 Wk 12 −20° C. 14.0 100.0 99.1 2-8°C. 14.0 100.1 99.1 25° C. 14.3 102.7 99.0 40° C. 13.7 98.2 98.9

The physical stability of the emulsion prepared in Example 8 was alsoevaluated. The physical stability was measured by the ability of theemulsion to maintain its average droplet size and the absence of largedroplet (>5 micron in diameter). The average droplet size is determinedby laser light scattering using a Malvern Zetasiziser 5000, and thepresence of large droplet (>5 micron in diameter) was examined byobserving the undiluted emulsion using an optical microscope at 400×magnifications. The physical stability data of the emulsion are providedin the table below:

Physical stability AVG Droplet Time Storage size Large droplets Visualpoint Condition (nm) (>5 micron in diameter) appearance 0 115 NoneUniform Wk 1 −20° C. 117 Many Uniform 2-8° C. 113 None Uniform 25° C.120 Some Uniform 40° C. 146 Some Uniform Wk 2 −20° C. 126 Many Uniform2-8° C. 115 None Uniform 25° C. 122 Some Uniform 40° C. 162 Many UniformWk 4 −20° C. 137 Many Uniform 2-8° C. 110 Some Uniform 25° C. 123 SomeUniform 40° C. 169 Some Uniform Wk 12 −20° C. 201 Many N/A 2-8° C. 116None Uniform 25° C. 143 Some Uniform 40° C. 117 Some Yellowish, viscous

Conclusion: Vinorelbine is chemically stable in the emulsion prepared inExample 8 at 2-8° C. or 25° C., and the emulsion is physically stable at2-8° C. for at least 12 weeks (3 months).

Example 10 Freeze-Drying Vinorelbine Emulsion

This study was to demonstrate the feasibility of converting a liquidemulsion to a freeze-dried emulsion or the “oil-in-solid dispersionsystem”, which is believed more stable than the liquid emulsion.

Both low pH (pH 3.75) and neutral pH (pH 7.14) emulsion formulationswere designed and prepared for freeze-drying or lyophilization study.

The formulation contained:

% (w/w) Vinorelbine bitartrate 1.39 Miglyol 812N 15 Soy lecithin 7.5Disodium EDTA dihydrate 0.005 Oleic acid 1.45 Sucrose 15 Deionized waterto QS 100

To prepare the freeze-dried emulsions, vinorelbine bitartrate, Miglyol812, phospholipon 90G and oleic acid were first dissolved in sufficientamount of dehydrated ethanol to form a clear solution. The ethanol wasremoved using a rotary evaporator under vacuum at room temperatureovernight to obtain an oil phase. The oil phase was mixed with anaqueous phase, which contained sucrose and sodium EDTA to form a crudeemulsion using a high shear homogenizer. The oleic acid used in thisformulation resulted in a low pH emulsion naturally. The crude emulsionwas then microfluidized for 6 passages to form a pH 3.75 final emulsion.

A portion of the pH 3.75 crude emulsion was adjusted to pH 7.14 using0.5N sodium hydroxide. The crude emulsion was then microfluidized for 6passages to form a pH 7.14 final emulsion. Both emulsions were filteredthrough 0.2-micron filters and filled into as 1 mL each in 5 mL vials or0.2 mL each in 2 mL vials. The height of the fill was about 3-4 mm. Thevials were partially stoppered with lyophile stoppers and freeze-driedusing a freeze-dryer (Dura-Stop™ mp by FTS System).

At the completion of the freeze-drying cycle, the freeze-dryer chamberwas back filled with nitrogen gas NF to about 95% of atmosphericpressure and then fully stoppered by collapsing the shelves. Thestoppered vials were sealed with aluminum crimp seals.

The dried emulsion or the oil-in-solid dispersion system was white“cakes” with uniform appearance. Prior to testing, the lyophile wasreconstituted with deionized water and mixed for 1-2 minutes to re-formthe liquid emulsion. The appearance (gross and microscopic) was recordedand the droplet size was determined (Table 9).

TABLE 9 VINORELBINE EMULSIONS PREPARED BY RECONSTITUTION OF THEOIL-IN-SOLID DISPERSION SYSTEM WITH DEIONIZED WATER Particle sizeEmulsion pH Appearance Microscopic appearance (nm) 3.75 Uniform No ≧5micron droplets 120.1 ± 3.0 7.14 Not uniform A lot of ≧5 micron dropletsNA

Conclusion: The pH 3.7 vinorelbine emulsion may be freeze-dried to formthe oil-in-solid dispersion system, and such oil-in-solid dispersionsystem can form an oil-in-water emulsion with size characteristicssimilar to the initial emulsion, upon dilution in water.

Example 11 Vein Irritation Test

The objective of this test is to compare vein irritation of avinorelbine emulsion of the present invention with a marketedvinorelbine solution product. The vinorelbine emulsion used was preparedas in Example 8 (without the freeze-drying step). The marketedvinorelbine solution (MINNUOBIN® marketed by Sino-Sanofi in China)contains 1% vinorelbine tartrate (equivalent to 1% vinorelbine freebase)in water at pH 3.5. This product has the same composition as Navelbine®,which is marketed in the U.S. by GlaxoSmithKline.

Six white rabbits were divided into three groups (two for each group,one male and one female). Each rabbit received daily an intravenousbolus injection through the marginal ear veins consecutively for 5 days.To Group I, 5% dextrose solution (D5W) was injected daily as a negativecontrol.

To Group II, MINNUOBIN® was administered at a dose of 1.68 mg/kg/dayafter dilution to 0.3% vinorelbine freebases in D5W as a positivecontrol.

To Group III, the vinorelbine emulsion prepared as in Example 8 wasadministered at a dose of 1.68 mg/kg/day either at 1% (undiluted) orafter dilution to 0.3% vinorelbine freebase in D5W.

In Group III treated with the vinorelbine emulsion prepared as inExample 8, no drug-related signs of vein irritation were observed byappearance examination and pathology histology. All observed changeswere due to mechanical punctuation during injection. Rabbits in Group II(treated with MINNUOBIN®) exhibited signs of mild to medium level veinirritation.

Conclusion: The vinorelbine emulsion of this invention did not causevein irritation, while the solution formulation resulted in significantvein irritation at the same intravenous dose.

Example 12 Acute Toxicity Test

The objective of this test is to compare acute toxicity of a vinorelbineemulsion prepared as in Example 8 with a marketed vinorelbine solutionproduct (MINNUOBIN®) in mice.

Mice (100) were divided into five (5) dose groups with 20 animals ineach group (10 males and 10 females). MINNUOBIN® and the vinorelbineemulsion as described in Example 8 were administered intravenously.Immediate reactions were observed and acute toxicity was calculatedusing the Bliss method.

The iv×1 acute toxicity LD50 values were calculated as:

Vinorelbine solution (MINNUOBIN ®): 37.56 (34.24-41.21) mg/kgVinorelbine emulsion (Example 8): 40.93 (37.75-44.38) mg/kg

Conclusion: There was no statistical difference between these twogroups.

All of the above U.S. patents, U.S. patent application publications,U.S. patent applications, foreign patents, foreign patent applicationsand non-patent publications referred to in this specification and/orlisted in the Application Data Sheet, are incorporated herein byreference, in their entirety.

From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1-20. (canceled)
 21. A method for treating cancer while reducing veinirritation, said method comprising administering a compositioncomprising a triglyceride oil, an emulsifier, a stabilizer, water, and ahighly water soluble drug, wherein (a) the composition is an emulsionhaving an oil and an aqueous phase, and (b) the drug is substantially inthe oil phase, to treat cancer.
 22. The method of claim 21, wherein theemulsion is at a pH range of 3 to
 5. 23. The method of claim 21, whereinthe drug alone is venous toxic.
 24. The method of claim 21, wherein thedrug is weakly basic.
 25. The method of claim 21, wherein the drug isselected from the group consisting of dopamine, ciprofloxacin,vancomycin, norvancomycin, doxorubicin, daunorubicin, vinca alkaloids,and pharmaceutically acceptable salts thereof.
 26. The method of claim21, wherein said drug is vinorelbine bitartrate having the chemical nameof 3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)] and the followingstructure: or another pharmaceutically acceptable salt of vinorelbine.27. The method of claim 21, wherein the triglyceride oil is atriglyceride having long chain fatty acids, a triglyceride having mediumchain fatty acids, or a mixture thereof.
 28. The method of claim 21,wherein the emulsifier is egg lecithin, soy lecithin, a syntheticphospholipid, or a mixture thereof.
 29. The method of claim 21, whereinthe stabilizer is a fatty acid, riboflavin-5-phosphate, vitamin-Esuccinate, cholesterol sulfate, or a mixture thereof.
 30. The method ofclaim 29, wherein the fatty acid is oleic acid or a pharmaceuticallyacceptable salt thereof.
 31. The method of claim 21, wherein the drughas an aqueous solubility of over 50 mg/ml.
 32. The method of claim 21,wherein the drug has an aqueous solubility of over 100 mg/ml.
 33. Themethod of claim 21, wherein the drug has an aqueous solubility of over500 mg/ml.
 34. The method of claim 21, wherein the drug has an aqueoussolubility of over 1000 mg/ml.
 35. The method of claim 21, wherein thecharge ratio of the drug to the stabilizer is 1:1 to 1:10.
 36. Themethod of claim 21, wherein no less than 90% of the drug is present inthe oil phase of the emulsion.
 37. The method of claim 21, wherein noless than 95% of the drug is present in the oil phase of the emulsion.38. The method of claim 21, wherein the drug in the composition is in aconcentration range of 1 to 50 mg/mL.